Reproducibility of double agar gel immunodiffusion test using stored serum and plasma from paracoccidioidomycosis patients

Background: Serological evaluation performed by double agar gel immunodiffusion test (DID) is used for diagnosis, evaluation of severity, management of paracoccidioidomycosis patients, and development of new clinical studies. For these reasons, the Botucatu Medical School of UNESP maintains a serum bank at the Experimental Research Unit with patient clinical data. This study aimed to evaluate the influence of the freeze-thaw cycle and different blood matrices on the titration of circulating antibodies. Methods: The study included 207 patients with confirmed (etiology-demonstrated) or probable (serology-demonstrated) paracoccidioidomycosis, and DID was performed with culture filtrate from Paracoccidioides brasiliensis B339 as antigen. First experiment: the antibody levels were determined in serum samples from 160 patients with the chronic form and 20 with the acute/subacute form, stored at ­80o C for more than six months. Second experiment: titers of 81 samples of serum and plasma with ethylenediaminetetraacetic acid (EDTA) or heparin, from 27 patients, were compared according to matrix and effect of storage at ­20o C for up to six months. Differences of titers higher than one dilution were considered discordant. Results: First experiment: test and retest presented concordant results in serum stored for up to three years, and discordant titers in low incidence in storage for four to six years but high incidence when stored for more than six years, including conversion from reagent test to non-reagent retest. Second experiment: serum, plasma-EDTA and plasma-heparin samples showed concordant titers, presenting direct correlation, with no interference of storage for up to six months. Conclusions: Storage at ­80o C for up to six years has no or little influence on the serum titers determined by DID, permitting its safe use in studies depending on this parameter. The concordant titrations in different blood matrices demonstrated that the plasma can be used for immunodiffusion test in paracoccidioidomycosis, with stability for at least six months after storage at ­20o C.(AU)

Molecular Epidemiology of Plasmodium species in Conflicted Federally Administered Tribal Area (FATA) Pakistan

Abstract Military conflicts have been significant obstacles in detecting and treating infectious disease diseases due to the diminished public health infrastructure, resulting in malaria endemicity. A variety of violent and destructive incidents were experienced by FATA (Federally Administered Tribal Areas). It was a struggle to pursue an epidemiological analysis due to continuing conflict and Talibanization. Clinical isolates were collected from Bajaur, Mohmand, Khyber, Orakzai agencies from May 2017 to May 2018. For Giemsa staining, full blood EDTA blood samples have been collected from symptomatic participants. Malaria-positive microscopy isolates were spotted on filter papers for future Plasmodial molecular detection by nested polymerase chain reaction (nPCR) of small subunit ribosomal ribonucleic acid (ssrRNA) genes specific primers. Since reconfirming the nPCR, a malariometric study of 762 patients found 679 positive malaria cases. Plasmodium vivax was 523 (77%), Plasmodium falciparum 121 (18%), 35 (5%) were with mixed-species infection (P. vivax plus P. falciparum), and 83 were declared negative by PCR. Among the five agencies of FATA, Khyber agency has the highest malaria incidence (19%) with followed by P. vivax (19%) and P. falciparum (4.1%). In contrast, Kurram has about (14%), including (10.8%) P. vivax and (2.7%) P. falciparum cases, the lowest malaria epidemiology. Surprisingly, no significant differences in the distribution of mixed-species infection among all five agencies. P. falciparum and P. vivax were two prevalent FATA malaria species in Pakistans war-torn area. To overcome this rising incidence of malaria, this study recommends that initiating malaria awareness campaigns in school should be supported by public health agencies and malaria-related education locally, targeting children and parents alike.

Resumo Os conflitos militares têm sido obstáculos significativos na detecção e tratamento de doenças infecciosas devido à diminuição da infraestrutura de saúde pública, resultando na endemicidade da malária. Uma variedade de incidentes violentos e destrutivos foi vivida pelas FATA (áreas tribais administradas pelo governo federal). Foi uma luta buscar uma análise epidemiológica devido ao conflito contínuo e à talibanização. Isolados clínicos foram coletados de agências Bajaur, Mohmand, Khyber e Orakzai, de maio de 2017 a maio de 2018. Para a coloração de Giemsa, amostras de sangue completo com EDTA foram coletadas de participantes sintomáticos. Isolados de microscopia positivos para malária foram colocados em papéis de filtro para futura detecção molecular plasmódica por reação em cadeia da polimerase aninhada (nPCR) de primers específicos de genes de subunidade ribossômica de ácido ribonucleico (ssrRNA). Desde a reconfirmação do nPCR, um estudo malariométrico de 762 pacientes encontrou 679 casos positivos de malária. Plasmodium vivax foi 523 (77%), Plasmodium falciparum 121 (18%), 35 (5%) eram com infecção de espécies mistas (P. vivax mais P. falciparum) e 83 foram declarados negativos por PCR. Entre as cinco agências da FATA, a agência Khyber tem a maior incidência de malária (19%), seguida por P. vivax (19%) e P. falciparum (4,1%). Em contraste, Kurram tem cerca de 14%, incluindo 10,8% casos de P. vivax e 2,7% P. falciparum, a epidemiologia de malária mais baixa. Surpreendentemente, não há diferenças significativas na distribuição da infecção de espécies mistas entre todas as cinco agências. P. falciparum e P. vivax foram duas espécies prevalentes de malária FATA na área devastada pela guerra no Paquistão. Para superar essa incidência crescente de malária, este estudo recomenda que o início de campanhas de conscientização sobre a malária na escola deve ser apoiado por agências de saúde pública e educação relacionada com a malária localmente, visando crianças e pais.

Morphometric analysis on erythrocytes of various livestock being reared in the Cholistan desert / Análise morfométrica de eritrócitos de vários pecuários criados no deserto de Colistão

The present study aimed to assess the influence of physiological exposure to erythrocyte indices; the morphometric parameters of the size of erythrocytes, as well as synergistic effects on the erythrocyte spectrum changes in various species being reared in different strip-wasteland conditions. We collected samples from 400 animals from the jugular veins and made 2000 blood smears, sample blood was taken by an expert veterinary doctor and transferred into an EDTA vacutainer. Blood smears were stained through the Pappenheim procedure and slides were photomicrographed under 100x objective. The parameters of morphometry of erythrocytes were largest in cattle after buffalo, sheep, while goats have the lowest one. The influence of age was significant p<0.05. The influence of sex was not significant in Cattle p<0.01, values of a parameter of erythrocytes were higher in young than in adults in sheep, goats, cattle, and buffalo. The size of erythrocytes in males was higher than females in sheep, goat, and buffalo but in cattle values of female erythrocytes were higher than in males. Further, gender, altitude, and age have a profound influence on the morphometric attributes of erythrocytes. It will be helpful in interpreting etiopathogenetic conditions in human beings and other animals.

O presente estudo teve como objetivo avaliar a influência da exposição fisiológica nos índices eritrocitários; os parâmetros morfométricos do tamanho dos eritrócitos, bem como os efeitos sinérgicos sobre as alterações do espectro eritrocitário em várias espécies criadas em diferentes condições de terreno baldio. Coletamos amostras de 400 animais das veias jugulares e fizemos 2.000 esfregaços de sangue, a amostra de sangue foi colhida por um médico veterinário especialista e transferida para um vacutainer de EDTA. Esfregaços de sangue foram corados pelo procedimento de Pappenheim e as lâminas foram fotomicrografadas sob objetiva de 100x. Os parâmetros de morfometria dos eritrócitos foram maiores em bovinos após bubalinos, ovinos, enquanto os caprinos apresentaram os menores. A influência da idade foi significativa p<0,05. A influência do sexo não foi significativa em Bovinos p<0,01, os valores de um parâmetro de eritrócitos foram maiores em jovens do que em adultos em ovinos, caprinos, bovinos e bubalinos. O tamanho dos eritrócitos nos machos foi maior que nas fêmeas em ovinos, caprinos e búfalos, mas em bovinos os valores de eritrócitos femininos foram maiores do que nos machos. Além disso, sexo, altitude e idade têm uma profunda influência nos atributos morfométricos dos eritrócitos. Será útil na interpretação de condições etiopatogenéticas em seres humanos e outros animais.

Influence on clinical biochemistry values of black-tufted marmosets (Callithrix penicillata) anesthetized with isoflurane or sevoflurane / Influência sobre valores de bioquímica clínica de saguis-de-tufo-preto (Callithrix penicillata) anestesiados com isoflurano ou sevoflurano

The objective of this study was to evaluate the clinical biochemistry behavior of Black-Tufted Marmosets (Callithrix penicillatta) submitted to blood collection without sedation and after general anesthesia with anesthetics isoflurane or sevoflurane. Blood collections were performed on (M1) day before anesthesia by physical restraint, and (M2) after anesthesia. There were four groups: Isoflurane (GI) and Sevoflurane (GS) using an anesthetic box. GIM: isoflurane induction with mask for a shorter period. Control group (GP): physical restraint in both moments. Plasma was separated and frozen to measure clinic biochemistry values. Urea was higher at M2 in groups GI and GP. AST was higher in M2 in GI, GS, and GP and only GI showed an increase in CK in M2. Glucose was higher in M1 in the GI, GS, and GP groups and fructosamine was higher in M2 in the GI. Stress caused by physical restraint can cause biochemical changes and these must be considered when interpreting the exams. Both the inhalational anesthetic isoflurane and sevoflurane did not cause clinically significant changes in clinical biochemistry results.

O objetivo desse estudo foi avaliar o comportamento da bioquímica clínica de saguis-do-tufo-preto (Callithrix penicillatta) submetidos à coleta sanguínea sem sedação e após anestesia geral, com os anestésicos isoflurano ou sevoflurano. As coletas de sangue foram realizadas: (M1) dia antes da anestesia por contenção física e (M2) após anestesia. Foram definidos quatro grupos: isoflurano (GI) e sevoflurano (GS), utilizando caixa anestésica; GIM: indução com isoflurano com máscara por um período menor; grupo controle (GCF): contenção física em ambos os momentos. O plasma com EDTA foi separado e congelado para realizar dosagem da bioquímica clínica. A ureia foi maior no M2 nos grupos GI e GCF. A AST foi maior no M2 nos GI, GS e GCF e somente o GI apresentou aumento de CK no M2. A glicose foi maior no M1 nos grupos GI, GS e GCF, e a frutosamina foi maior no M2 no GI. O estresse causado pela contenção física pode causar alterações bioquímicas e essas devem ser levadas em consideração na interpretação dos exames. Tanto o anestésico inalatório isoflurano quanto o sevoflurano não causaram alterações clinicamente significantes nos resultados da bioquímica clínica.

Molecular Epidemiology of Plasmodium species in Conflicted Federally Administered Tribal Area (FATA) Pakistan / Epidemiologia molecular de espécies de plasmódio em área tribal sob administração federal em conflito do Paquistão

Military conflicts have been significant obstacles in detecting and treating infectious disease diseases due to the diminished public health infrastructure, resulting in malaria endemicity. A variety of violent and destructive incidents were experienced by FATA (Federally Administered Tribal Areas). It was a struggle to pursue an epidemiological analysis due to continuing conflict and Talibanization. Clinical isolates were collected from Bajaur, Mohmand, Khyber, Orakzai agencies from May 2017 to May 2018. For Giemsa staining, full blood EDTA blood samples have been collected from symptomatic participants. Malaria-positive microscopy isolates were spotted on filter papers for future Plasmodial molecular detection by nested polymerase chain reaction (nPCR) of small subunit ribosomal ribonucleic acid (ssrRNA) genes specific primers. Since reconfirming the nPCR, a malariometric study of 762 patients found 679 positive malaria cases. Plasmodium vivax was 523 (77%), Plasmodium falciparum 121 (18%), 35 (5%) were with mixed-species infection (P. vivax plus P. falciparum), and 83 were declared negative by PCR. Among the five agencies of FATA, Khyber agency has the highest malaria incidence (19%) with followed by P. vivax (19%) and P. falciparum (4.1%). In contrast, Kurram has about (14%), including (10.8%) P. vivax and (2.7%) P. falciparum cases, the lowest malaria epidemiology. Surprisingly, no significant differences in the distribution of mixed-species infection among all five agencies. P. falciparum and P. vivax were two prevalent FATA malaria species in Pakistan's war-torn area. To overcome this rising incidence of malaria, this study recommends that initiating malaria awareness campaigns in school should be supported by public health agencies and malaria related education locally, targeting children and parents alike.

Os conflitos militares têm sido obstáculos significativos na detecção e tratamento de doenças infecciosas devido à diminuição da infraestrutura de saúde pública, resultando na endemicidade da malária. Uma variedade de incidentes violentos e destrutivos foi vivida pelas FATA (áreas tribais administradas pelo governo federal). Foi uma luta busca ruma análise epidemiológica devido ao conflito contínuo e à talibanização. Isolados clínicos foram coletados de agências Bajaur, Mohmand, Khyber e Orakzai, de maio de 2017 a maio de 2018. Para a coloração de Giemsa, amostras de sangue completo com EDTA foram coletadas de participantes sintomáticos. Isolados de microscopia positivos para malária foram colocados em papéis de filtro para futura detecção molecular plasmódica por reação em cadeia da polimerase aninhada (nPCR) de primers específicos de genes de subunidade ribossômica de ácido ribonucleico (ssrRNA). Desde a reconfirmação do nPCR, um estudo malariométrico de 762 pacientes encontrou 679 casos positivos de malária. Plasmodium vivax foi 523 (77%), Plasmodium falciparum 121 (18%), 35 (5%) eram com infecção de espécies mistas (P. vivax mais P. falciparum) e 83 foram declarados negativos por PCR. Entre as cinco agências da FATA, a agência Khyber tem a maior incidência de malária (19%), seguida por P. vivax (19%) e P. falciparum (4,1%). Em contraste, Kurram tem cerca de 14%, incluindo 10,8% casos de P. vivax e 2,7% P. falciparum, a epidemiologia de malária mais baixa. Surpreendentemente, não há diferenças significativas na distribuição da infecção de espécies mistas entre todas as cinco agências. P. falciparum e P. vivax foram duas espécies prevalentes de malária FATA na área devastada pela guerra no Paquistão. Para superar essa incidência crescente de malária, este estudo recomenda que o início de campanhas de conscientização sobre a malária na escola deve ser apoiado por agências de saúde pública e educação relacionada com a malária localmente, visando crianças e pais.

Molecular Epidemiology of Plasmodium species in Conflicted Federally Administered Tribal Area (FATA) Pakistan / Epidemiologia molecular de espécies de plasmódio em área tribal sob administração federal em conflito do Paquistão

Military conflicts have been significant obstacles in detecting and treating infectious disease diseases due to the diminished public health infrastructure, resulting in malaria endemicity. A variety of violent and destructive incidents were experienced by FATA (Federally Administered Tribal Areas). It was a struggle to pursue an epidemiological analysis due to continuing conflict and Talibanization. Clinical isolates were collected from Bajaur, Mohmand, Khyber, Orakzai agencies from May 2017 to May 2018. For Giemsa staining, full blood EDTA blood samples have been collected from symptomatic participants. Malaria-positive microscopy isolates were spotted on filter papers for future Plasmodial molecular detection by nested polymerase chain reaction (nPCR) of small subunit ribosomal ribonucleic acid (ssrRNA) genes specific primers. Since reconfirming the nPCR, a malariometric study of 762 patients found 679 positive malaria cases. Plasmodium vivax was 523 (77%), Plasmodium falciparum 121 (18%), 35 (5%) were with mixed-species infection (P. vivax plus P. falciparum), and 83 were declared negative by PCR. Among the five agencies of FATA, Khyber agency has the highest malaria incidence (19%) with followed by P. vivax (19%) and P. falciparum (4.1%). In contrast, Kurram has about (14%), including (10.8%) P. vivax and (2.7%) P. falciparum cases, the lowest malaria epidemiology. Surprisingly, no significant differences in the distribution of mixed-species infection among all five agencies. P. falciparum and P. vivax were two prevalent FATA malaria species in Pakistan's war-torn area. To overcome this rising incidence of malaria, this study recommends that initiating malaria awareness campaigns in school should be supported by public health agencies and malaria related education locally, targeting children and parents alike.(AU)

Os conflitos militares têm sido obstáculos significativos na detecção e tratamento de doenças infecciosas devido à diminuição da infraestrutura de saúde pública, resultando na endemicidade da malária. Uma variedade de incidentes violentos e destrutivos foi vivida pelas FATA (áreas tribais administradas pelo governo federal). Foi uma luta busca ruma análise epidemiológica devido ao conflito contínuo e à talibanização. Isolados clínicos foram coletados de agências Bajaur, Mohmand, Khyber e Orakzai, de maio de 2017 a maio de 2018. Para a coloração de Giemsa, amostras de sangue completo com EDTA foram coletadas de participantes sintomáticos. Isolados de microscopia positivos para malária foram colocados em papéis de filtro para futura detecção molecular plasmódica por reação em cadeia da polimerase aninhada (nPCR) de primers específicos de genes de subunidade ribossômica de ácido ribonucleico (ssrRNA). Desde a reconfirmação do nPCR, um estudo malariométrico de 762 pacientes encontrou 679 casos positivos de malária. Plasmodium vivax foi 523 (77%), Plasmodium falciparum 121 (18%), 35 (5%) eram com infecção de espécies mistas (P. vivax mais P. falciparum) e 83 foram declarados negativos por PCR. Entre as cinco agências da FATA, a agência Khyber tem a maior incidência de malária (19%), seguida por P. vivax (19%) e P. falciparum (4,1%). Em contraste, Kurram tem cerca de 14%, incluindo 10,8% casos de P. vivax e 2,7% P. falciparum, a epidemiologia de malária mais baixa. Surpreendentemente, não há diferenças significativas na distribuição da infecção de espécies mistas entre todas as cinco agências. P. falciparum e P. vivax foram duas espécies prevalentes de malária FATA na área devastada pela guerra no Paquistão. Para superar essa incidência crescente de malária, este estudo recomenda que o início de campanhas de conscientização sobre a malária na escola deve ser apoiado por agências de saúde pública e educação relacionada com a malária localmente, visando crianças e pais.(AU)

Comparative effectiveness of EDTA and citric acid assisted phytoremediation of Ni contaminated soil by using canola (Brassica napus) / Eficácia comparativa da fitorremediação assistida por EDTA e ácido cítrico de solo contaminado com Ni, usando canola (Brassica napus)

Phytoremediation is an ecofriendly technique to clean heavy metals from contaminated soil by the use of high biomass producing plant species. Chelators can help to improve this biological technique by increasing metal solubility. Therefore, a pot experiment was conducted to determine the effect of the chelators EDTA and citric acid (CA) in phytoremediation of Ni contaminated soil by using Brassica napus (canola). Two cultivars of B. napus, Con-II (tolerant) and Oscar (sensitive), were selected after screening and exposed to NiSO4 at 30 ppm at the time of sowing. CA (10 mM) and EDTA (1.5 mM) were applied either alone or in combination with each other after two weeks of Ni treatments. Different parameters like morpho-physiological and biochemical data were recorded after 15 days of chelate application. The results highlighted the successful use of chelating agents (CA and EDTA) not only to ameliorate Ni stress but also to enhance Ni accumulation which is prerequisite for phytoremediation. The basal application of 10 mMCA and 1.5 mM EDTA concentration proved to be effective for the growth of plants. The combination of chelating agents failed to show any synergistic effects.(AU)

A fitorremediação é uma técnica ecologicamente correta para limpar metais pesados ​​de solo contaminado pelo uso de espécies vegetais produtoras de alta biomassa. Os quelantes podem ajudar a melhorar esta técnica biológica aumentando a solubilidade do metal. Para tanto, foi realizado um experimento em vaso para determinar o efeito dos quelantes EDTA e ácido cítrico (AC) na fitorremediação de solo contaminado com Ni, utilizando Brassica napus (canola). Duas cultivares de B. napus, Con-II (tolerante) e Oscar (sensível) foram selecionadas após triagem e expostas a NiSO4 a 30 ppm no momento da semeadura. CA (10 mM) e EDTA (1,5 mM) foram aplicados sozinhos ou em combinação um com o outro após duas semanas de tratamentos com Ni. Diferentes parâmetros como dados morfofisiológicos e bioquímicos foram registrados após 15 dias de aplicação de quelato. Os resultados destacaram o uso bem-sucedido de agentes quelantes (CA e EDTA) não apenas para melhorar o estresse de Ni, mas também para aumentar o acúmulo de Ni, um pré-requisito para a fitorremediação. A aplicação basal da concentração de 10 mMCA e 1,5 mM de EDTA mostrou-se eficaz para o crescimento das plantas. A combinação de agentes quelantes não mostrou quaisquer efeitos sinérgicos.(AU)

An analytical method for the quantitative determination of iron ion chelating capacity: development and validation

Iron is a fundamental microelement for human life; however, deficiencies or excesses of these metal ions can cause severe complications and mortality. Chelators are compounds that bind and inhibit iron. Ultraviolet-visible (UV-vis) spectrophotometric methods are key analytical tools in the identification of chemical entities, with the benefits of having good precision and accuracy, and the equipment being easily available as well as quick and simple to implement. In this study, we aimed to provide an alternative, cheaper method for the quantification of iron ion chelation by substituting ferrozine for gallic acid and validating its use with UV-vis according to official ANVISA and ICH guidelines. The parameters assessed were specificity, linearity, precision, accuracy, robustness, and finally, the percentage of iron ions chelating was calculated. The results demonstrated that this method was accurate, simple, specific, selective, precise, and reproducible, and was successfully validated for the determination of iron ions chelating. The percentage of iron ions chelating, promoted by the standard chelator EDTA, was 45% and 47% for Fe2+ and Fe3+, respectively. It is concluded that this new method is beneficial in terms of its simplicity, rapidness, low cost, and the fact that it produces very low levels of dangerous residues.(AU)

Bio-efficacy of iron and zinc fortified wheat flour along with bio-assessment of its hepatic and renal toxic potential / Bioeficácia da farinha de trigo fortificada com ferro e zinco e bioavaliação do seu potencial tóxico hepático e renal

Study was planned to assess the bio-efficiency along with toxicity of iron and zinc fortified whole wheat flour in Sprague dawley albino rats. Whole wheat flour was fortified with different dosage of sodium iron EDTA (NaFeEDTA), ferrous sulphate (FeSO4), zinc oxide (ZnO) and zinc sulphate (ZnSO4). The rats (n=3) in each group were fed on fortified wheat flour for 2 months. Liver biomarkers including alkaline phosphatase (ALP), alanine transaminase (ALT), aspartate aminotransferase (AST) and bilirubin were recorded from serum samples. Increased concentration of ZnSO4 affected the liver biomarkers to be highest among all whereas, bilirubin levels were less than the rats fed on control diet. The above mentioned fortificants have negligible effect on renal biomarkers including creatinine and urea. Moreover, hematological parameters were also checked and reportedly, sodium iron EDTA fed rats presented highest amount of hemoglobin, iron and total iron binding capacity. Highest zinc level was observed in rats fed on whole wheat flour fortified with 60mg/Kg Zinc oxide. Microscopic observation of liver tissue depicted that rats fed on iron and zinc fortified wheat flour have more toxic effects whereas, histopathology presentation of kidney tissue has least toxic impact. It has been concluded that mandatory fortification of wheat flour with iron and zinc may cause increased serum biomarkers along with toxicity of vital organs like liver, hence fraction of wheat flour may be fortified to fulfill the requirements of deprived and vulnerable group.

O estudo foi planejado para avaliar a bioeficiência juntamente com a toxicidade da farinha de trigo integral fortificada com ferro e zinco em ratos albinos Sprague dawley. A farinha de trigo integral foi fortificada com diferentes dosagens de ferro sódico EDTA (NaFeEDTA), sulfato ferroso (FeSO4), óxido de zinco (ZnO) e sulfato de zinco (ZnSO4). Os ratos (n = 3) de cada grupo foram alimentados com farinha de trigo fortificada por 2 meses. Biomarcadores hepáticos incluindo fosfatase alcalina (ALP), alanina transaminase (ALT), aspartato aminotransferase (AST) e bilirrubina foram registrados a partir de amostras de soro. O aumento da concentração de ZnSO4 afetou os biomarcadores hepáticos como sendo os mais altos entre todos, enquanto os níveis de bilirrubina foram menores do que os ratos alimentados com dieta controle. Os fortificantes mencionados acima têm efeito insignificante nos biomarcadores renais, incluindo creatinina e ureia. Além disso, os parâmetros hematológicos também foram verificados e, segundo relatos, os ratos alimentados com EDTA de sódio e ferro apresentaram maior quantidade de hemoglobina, ferro e capacidade total de ligação de ferro. O maior nível de zinco foi observado em ratos alimentados com farinha de trigo integral fortificada com 60 mg/Kg de óxido de zinco. A observação microscópica do tecido hepático mostrou que ratos alimentados com farinha de trigo fortificada com ferro e zinco têm mais efeitos tóxicos, enquanto a apresentação histopatológica do tecido renal tem menos impacto tóxico. Concluiu-se que a fortificação obrigatória da farinha de trigo com ferro e zinco pode causar aumento dos biomarcadores séricos juntamente com toxicidade de órgãos vitais como o fígado, portanto a fração da farinha de trigo pode ser fortificada para atender os requisitos do grupo carente e vulnerável.

Erythrogram and oxidative stress in confined cattle fed with Brachiaria sp hay and supplemented with antioxidants / Eritrograma e estresse oxidativo em bovinos confinados alimentados com feno de Brachiaria sp. e suplementados com antioxidantes

Brachiaria sp contains sporidesmin that can be oxidized by lipoperoxidation and cause oxidative stress. In the present study we evaluated the effects of different antioxidants on lipoperoxidation of erythrocytes from Nelore cattle fed with Brachiaria sp hay. The experimental design was entirely randomized, in which 40 whole male cattle were divided into five treatments (G1: control - no supplementation; G2: selenium and vitamin E supplementation; G3: zinc supplementation; G4: selenium supplementation and G5: vitamin E supplementation) and allocated in feedlot pens for 105 days. The samples heparinized and withethylenediaminetetraacetic acid (EDTA) were obtained every 28 days for hematological and oxidative stress evaluation (0, 28 56, 84 and 105 days). In the erythrogram total erythrocyte count, hemoglobin, and hematocrit (Ht) were measured. For the evaluation of oxidative stress, in order to analyze the characteristics of the erythrocyte membrane, the thiobarbituric acid reactive substances (TBARS), total glutathione (GSH-T), glutathione peroxidase (GSH-Px), catalase (CAT) and superoxide dismutase (SOD) were determined. The results showed that regardless of the treatment there was no oxidative stress during the experimental confinement period and that the joint association of selenium and vitamin E in the bovine diet provided a lower incidence of deleterious alterations on erythrocytes.

As Brachiaria sp contêm esporidesminas que podem ser oxidadas por lipoperoxidação e ocasionar estresse oxidativo. No presente estudo foram avaliados os efeitos de diferentes antioxidantes na lipoperoxidação dos eritrócitos de bovinos da raça Nelore, alimentados com feno de Brachiaria sp. O delineamento experimental foi inteiramente casualizado, em que 40 bovinos machos, inteiros, foram divididos, em cinco tratamentos (G1: controle - sem suplementação; G2: suplementação de selênio e vitamina E; G3: suplementação de zinco; G4: suplementação de selênio e G5: suplementação de vitamina E) e alocados em baias de confinamento, por 105 dias. As amostras de plasma heparinizado ou com ácido etilenodiamino tetra-acético (EDTA) foram obtidas a cada 28 dias para avaliação hematológica e de estresse oxidativo (0, 28 56, 84 e 105 dias). No eritrograma foi mensurado a contagem total de eritrócitos, a hemoglobina e o volume globular (VG). Para a avaliação do estresse oxidativo, com o objetivo de analisar as características da membrana do eritrócito foram determinadas as substâncias reativas ao ácido tiobarbitúrico (TBARS), glutationa total (GSH-T), glutationa peroxidase (GSH-Px), catalase (CAT) e superóxido dismutase (SOD). Os resultados demonstraram que independente do tratamento não houve estresse oxidativo durante o período do confinamento experimental e que a associação conjunta de selênio e vitamina E na dieta dos bovinos proporcionaram menor incidência de alterações deletérias sobre os eritrócitos.

Free-living capybaras (Hydrochoerus hydrochaeris) in an urban area in Brazil: biochemical and hematological parameters

Capybaras have found favorable conditions for survival and reproduction in green urban environments. In recent years, the population of these large rodents has been increasingly abundant in several brazilian cities such as Uberlândia, a municipality of the southeastern region with a Cerrado biome. Capybaras are important in the Brazilian Spotted Fever epidemiological chain, by amplifying infection rates of the vector population. However, knowledge of this host's physiology is scarce. Thus, the aim of this work was to describe hematological and biochemical parameters of free-living capybaras groups in urbanized areas in the city of Uberlândia, Minas Gerais State, Brazil. Capybaras were captured in 4 different locations of Uberlândia city, Minas Gerais state, including 1 Condominium (P1), 1 Private Market Garden (P2), 1 Private Club (P3) and 1 Municipal Park (P4). The animals were baited into an octagonal iron corral and chemically contained with anesthetic darts. After sedated, blood was collected from the femoral vein in tubes with and without EDTA. Biochemical evaluation, hematological analysis with differential leukocyte counts and search for Dirofilaria sp. were done. The blood count and biochemistry values obtained from animals of different ages, sex and sectors (P1, P2, P3 and P4) were submitted to the Shapiro-Wilk normality test, considering 95% significance. Values that had a normal distribution were subjected to ANOVA tests followed by Student's t-test. Values that did not follow normality were submitted to the Kruskal-Wallis test, to obtain a P-value, with a significance level of 95%. A total of 19 capybaras were captured: 4 in P1, 6 in P2, 4 in P3 and 5 in P4. From the 19 animals, 13 were females (68.42%) and 6 were males (31.57%), 12 adults (63.15%) and 7 juveniles (36.84%). Apart from occasional skin scars and moderate to intense Ambyomma spp. tick infestations, all captured animals were healthy on a broad examination. From 5 animals captured in P4, despite the use of anticoagulant, blood from 4 animals clotted fast. No microfilariae were found in the thick drop test in any of the 19 animals sampled, and in 2 adult female capybaras captured in P1, Kurloff cells were observed. Hematological and biochemical values presented no major differences when comparing sex and age. Nevertheless, differences in liver and kidney profile were observed between the capybara groups, including ALT, alkaline phosphorus, BUN and creatinine. Blood from 4 animals clotted fast, despite the use of EDTA tubes. Blood clotting of samples with anticoagulant in this work could be associated with some physiological features inherent to capybaras. Many attempts were required to obtain enough blood from each individual due to the rapid hemostasis, what come in accordance with reports in literature. Kurloff cells were observed in 2 adult female capybaras captured in P1, which can be found in peripheral blood of female rodents during follicular phase of estrous cycle. Hematological and biochemical values differences in liver enzymes such as ALT and alkaline phosphorus, and kidney profile enzymes including BUN and creatinine could be associated to capture stress or dietetic variation between groups. Despite statistical relevant, the values were still in accordance with other works, although comparisons should be done with caution since various environments exert a diverse array of stimulus upon the animals such as parasitic, infective, stress, nutritional, social and undoubtedly blood parameters mirror them. In conclusion, this work contributes to the standardization of free-living capybaras' physiological parameters in urban areas.(AU)

Follow-up of dairy cattle naturally infected by Trypanosoma vivax after treatment with isometamidium chloride / Acompanhamento de bovinos leiteiros naturalmente infectados com Trypanosoma vivax após tratamento com cloreto de isometamidium

Trypanosoma vivax infections cause nonspecific clinical signs in cattle associated with aparasitemic intervals, making disease diagnosis a challenge. In Brazil, diminazene aceturate and isometamidium chloride (ISM) are available to treat bovine trypanosomosis. The objective of this study was to follow-up, by molecular and serological techniques, dairy cattle naturally infected by T. vivax after ISM treatment. Thirty cattle naturally infected with T. vivax received two applications of ISM, at a dosage of 1.0 mg/kg intramuscularly, on days 0 and 150. For T. vivax diagnosis, EDTA-blood and serum samples were evaluated on 0, 7, 15, 30, 60, 90, 120, 150, 180, 210, and 240 days after treatment PCR, Loop-mediated isothermal amplification (LAMP) and ELISA. Animals with persistent detection of T. vivax DNA by both PCR and LAMP were found and continuous detection of anti-T. vivax IgG antibodies by ELISA, suggesting the presence of T. vivax resistance to ISM. The combination of LAMP and ELISA tests can prevent misdiagnosis of the parasite clearance in treated cattle, contributing to better disease control. This is the first experiment that demonstrates the persistence infection of T. vivax under ISM treatment in a natural infected herd and evidence of ISM chemotherapy-resistant T. vivax in Brazil.(AU)

Em bovinos, infecções por Trypanosoma vivax geram sinais clínicos inespecíficos que, associados a intervalos aparasitêmicos, faz com que o diagnóstico da enfermidade seja desafiador. No Brasil, somente aceturato de diaminazeno e cloridrato de isometamidum (ISM) estão disponíveis para o tratamento da tripanossomose bovina. Este trabalho teve como objetivo acompanhar bovinos leiteiros naturalmente infectados por T. vivax, após o tratamento com ISM por meio de técnicas moleculares e sorológica. Foram utilizados 30 bovinos naturalmente infectados com T. vivax, sendo estes tratados com duas aplicações de ISM, na dosagem de 1,0 mg/kg por via intramuscular profunda, nos dias 0 e 150. Foram avaliadas, para diagnóstico de T. vivax, amostras de sangue acrescido de EDTA e soro, colhidas nos 0, 7, 15, 30, 60, 90, 120, 150, 180, 210 e 240 dias após os tratamentos pela reação em cadeia da polimerase (PCR), amplificação circular isotérmica do DNA (LAMP) e ensaio de imunoabsorção enzimático (ELISA). Verificou-se a presença de animais com persistência na detecção de DNA de T. vivax pela PCR e LAMP, bem como detecção contínua de anticorpos IgG anti-T. vivax pelo método de ELISA, sugerindo a presença de resistência de T. vivax ao ISM. A combinação dos testes LAMP e ELISA pode evitar falsos diagnósticos da eliminação do parasita nos bovinos tratados, contribuindo para um melhor controle da doença. Este é o primeiro experimento que demonstra infecção persistente do T. vivax em rebanho naturalmente infectado, tratado com ISM, e evidencia possível resistência ao quimioterápico no Brasil.(AU)

How much does the platelet aggregate influence the total leukocyte count? Comparison between manual and automated impedance methods in domestic cats

Background: Automated hematology analyzers have been developed to optimize the time between analyses and havepromising precision and accuracy. Complete blood count (CBC) is often requested as part of veterinary clinical examination. Automated analyzers are often used to determine CBCs, since processing as well as container-related errors mayoccur owing to variable sizes, aggregates, white or red blood cell fragments, and effects of EDTA on cell morphology.Platelet aggregates frequently occur in felines, with studies reporting a prevalence of approximately 71%. The aim of thepresent study was to evaluate the influence of exercise aggregates on the global white blood cell count of domestic catsusing automated hematological counters with the impedance method.Materials, Methods & Results: Blood samples of 140 cats, irrespective of age, sex, and breed, were collected into EDTAcontaining tubes. The samples were obtained via routine clinical examinations at the Veterinary Hospital of the FederalRural University of Rio de Janeiro (UFRRJ) and processed at the Veterinary Parasitology Experimental ChemotherapyLaboratory (LQEPV), belonging to the same institution. All the samples were processed on the Sysmex pocH-100iVDiff automated hematology apparatus according to the manufacturer’s recommendations. Leukocyte counts were alsomanually determined using a duplicate Neubauer chamber. Standard dilutions were prepared immediately after theautomated analysis. To identify the occurrence of platelet aggregates, a blood smear was made and visualized undera brightfield microscope at a magnification of 10× and scored 0 to 3 (G1, G2, G3, and G4) based on the aggregationintensity. In case of changes, the groups were subdivided according to the intensity of occurrence. Of the 140 samples...

How much does the platelet aggregate influence the total leukocyte count? Comparison between manual and automated impedance methods in domestic cats

Background: Automated hematology analyzers have been developed to optimize the time between analyses and havepromising precision and accuracy. Complete blood count (CBC) is often requested as part of veterinary clinical examination. Automated analyzers are often used to determine CBCs, since processing as well as container-related errors mayoccur owing to variable sizes, aggregates, white or red blood cell fragments, and effects of EDTA on cell morphology.Platelet aggregates frequently occur in felines, with studies reporting a prevalence of approximately 71%. The aim of thepresent study was to evaluate the influence of exercise aggregates on the global white blood cell count of domestic catsusing automated hematological counters with the impedance method.Materials, Methods & Results: Blood samples of 140 cats, irrespective of age, sex, and breed, were collected into EDTAcontaining tubes. The samples were obtained via routine clinical examinations at the Veterinary Hospital of the FederalRural University of Rio de Janeiro (UFRRJ) and processed at the Veterinary Parasitology Experimental ChemotherapyLaboratory (LQEPV), belonging to the same institution. All the samples were processed on the Sysmex pocH-100iVDiff automated hematology apparatus according to the manufacturers recommendations. Leukocyte counts were alsomanually determined using a duplicate Neubauer chamber. Standard dilutions were prepared immediately after theautomated analysis. To identify the occurrence of platelet aggregates, a blood smear was made and visualized undera brightfield microscope at a magnification of 10× and scored 0 to 3 (G1, G2, G3, and G4) based on the aggregationintensity. In case of changes, the groups were subdivided according to the intensity of occurrence. Of the 140 samples...(AU)

Dermatite associada ao consumo de farelo de arroz desengordurado em bovinos na região sul do Estado do Rio Grande do Sul / Dermatitis associated with the intake of defatted rice bran by cattle in the Southern region of the state of Rio Grande do Sul

Background: Dermatitis associated with defatted rice bran (DRB) seems to be an underdiagnosed disease in Brazilian confined herds, characterized by localized skin lesions that develop mainly on hind limbs, and can affect any animal category. In this context, the goal of the present study was to describe an outbreak of dermatitis associated with the consumption of defatted rice bran on a property in the State of Rio Grande do Sul, demonstrating the clinical, hematological and epidemiological characteristics of the animals, as well as alternatives for the definitive diagnosis. Cases: Clinical and epidemiological data were obtained by anamnesis with the technician responsible for the property. Aberdeen Angus and crossbred males, with 24 to 36 months of age, with an average of 413 kg, from a property in the interior of the state of Rio Grande do Sul were analyzed. From a batch of 45 cattle, from 24 to 36 months of age, 20 presented lesions on hind limbs 8-17 days after supplementation of a commercial feed containing defatted rice bran. To assist in the diagnosis, blood samples were drawn into vacuum tubes with 10% EDTA, for a complete blood count with the investigation of hemoparasites, and without anticoagulant, for liver function tests. In addition, lesion tissue samples were also collected for bacteriological, mycological and histopathological examination and the ration offered to the animals, for intradermal tests. There were no significant hematological and biochemical changes in animals that developed DRB dermatitis, except when they have an associated secondary infection. The fungal research was negative. The bacterial culture revealed a growth of Staphylococcus aureus, possibly due to secondary infection resulting from the lesions. In histopathological examination, lesions were characterized by areas of alopecia, thickening of the epidermis, forming dry, thick crusts, and some ulcerative and serosanguineous lesions. Histologically, lesions were characterized by marked hyperkeratosis, ulceration, and in the superficial dermis, intense inflammatory infiltrate of eosinophils and lymphocytes. Histologic changes, although not pathognomonic, are typically described in this disease. The intradermal test was performed to contribute to the diagnosis of the disease, where a significant increase in volume was found between measurements on animals that developed the disease. It is believed that the disease is produced due to a food hypersensitivity as a consequence of the high protein level in DRB. The results of the intradermal test indicate that the animals developed hypersensitivity and reaction to proteins, and further research is required to determine the protein fraction leading to hypersensitivity reactions. Discussion: In the present study, acute lesions in hind limbs in a significant number of animals of the same batch in a short period of time after supplementation with a diet containing defatted rice bran, enabled a clinical diagnosis suggestive of dermatitis associated with DRB consumption. Through epidemiological data, reactive intradermal test, associated with the findings of the histopathological exam, which showed characteristic lesions of the disease (alopecia, erythema, epidermis thickening, with the formation of thick crusts, usually on hind limbs in the region of the hoof coronary band, progressing to pastern and fetlock), it was possible to establish the clinical-pathological diagnosis of dermatitis associated with the consumption of defatted rice bran.

Effects of chelating calcium in cryopreservation extender on frozen-thawed dog semen / [Efeitos da quelação de cálcio no meio de criopreservação no sêmen descongelado de cão]

We evaluated the effect of reducing free calcium in the cryopreservation medium, using the calcium chelator ethylene diamine tetracetic acid (EDTA) at 0.3% and 0.5% concentrations. Three male mixed breed dogs were subjected to semen collection by digital manipulation (n=16). Each ejaculate was divided in three aliquots, and each one was diluted in TRIS-glucose-egg yolk extender with 6% glycerol and 0.5% Equex STM Paste® (TGE, control); and added with 0.3% EDTA (EDTA 0.3) or 0.5% EDTA (EDTA 0.5). Calcium concentration reduced in EDTA 0.3 and all the calcium ions were chelated in EDTA 0.5. The EDTA addition did not affect sperm morphology or plasma membrane integrity; however, by removing all free calcium (EDTA 0.5), the sperm motility reduced (64.7% in TGE and 45% in EDTA 0.5; p<0.05). Acrosome integrity and sperm binding ability were not improved by calcium chelation. The failure to prevent the premature AR may explain why sperm longevity was not affected by calcium removal. Thus, the partial or complete calcium removal, through EDTA addition, is not able to prevent acrosomal damage or premature acrosomal reaction, and therefore does not improve the dog sperm binding ability.(AU)

Avaliou-se o efeito da redução do cálcio livre no meio de congelamento, usando-se o quelante de cálcio etilenodiaminotetracético (EDTA) a 0,3% e 0,5%. Três cães machos sem raça definida foram submetidos à coleta de sêmen por manipulação digital (n=16). Cada ejaculado foi diluído em diluidor controle com TRIS-glicose - gema de ovo (TGE, controle), ou em diluidor TGE enriquecido com 0,3% (EDTA 0,3) ou 0,5% de EDTA (EDTA 0,5). A concentração de cálcio reduziu no meio EDTA 0,3, e todos os íons de cálcio foram quelados no meio EDTA 0,5. A adição do EDTA e a consequente quelação do cálcio não afetaram a morfologia espermática ou a integridade da membrana plasmática, no entanto, ao remover todo o cálcio do meio (EDTA 0,5), a motilidade espermática se reduziu (64,7% no TGE e 45% no EDTA 0,5; P<0,05). A integridade do acrossoma e a capacidade de ligação do espermatozoide não melhoraram com a quelação do cálcio. Apesar da influência da concentração de cálcio sobre a motilidade espermática após o descongelamento, a falha em prever a reação acrossomal prematura pode explicar por que a longevidade espermática não foi afetada pela remoção do cálcio no meio. Dessa forma, a remoção parcial ou total do cálcio, por meio da adição de EDTA, não é capaz de prevenir o dano no acrossoma ou a reação acrossomal prematura e, portanto, não aumenta a capacidade do espermatozoide de se ligar ao oócito.(AU)

Partial characterization of digestive proteases in juveniles of Microphis brachyurus (short-tailed pipefish) (Syngnathiformes: Syngnathidae)

RESUMEN El pez pipa (Microphis brachyurus) es un organismo dulceacuícola con alto potencial económico para la acuarofilia; sin embargo, es necesario implementar su cultivo a través de estudios de fisiología digestiva. Se evaluó el efecto del pH, temperatura e inhibidores sobre las actividades enzimáticas de proteasas ácidas y alcalinas. El pH óptimo de proteasas estomacales es de 2, mientras que el de proteases intestinales es de 10. La temperatura óptima de proteasas ácidas es de 35 ºC y las alcalinas de 45 ºC. La estabilidad térmica para proteasas ácidas y alcalinas es a los 35 ºC (más de 100% de actividad residual). La estabilidad a los diferentes pH de las proteasas ácidas es en 2 (50 % de la actividad residual), mientras que para las proteasas alcalinas es en 10 (90 % de la actividad residual). Las proteasas ácidas fueron inhibidas en 80% con pepstatina A y las proteasas alcalinas fueron altamente inhibidas con TLCK para tripsina (75%) y TPCK quimitripsina (80%). Finalmente, las metaloproteasas fueron inactivadas con EDTA en 70%. En conclusión, M. brachyurus tiene una buena capacidad digestiva al degradar una amplia variedad de proteinas debido a su alta actividad proteolítica.

ABSTRACT Short-tailed pipe fish (Microphis brachyurus) is a freshwater organism with high economic potential for the aquarium hobby, so it is necessary to implement methods to promote its culture through studies of digestive physiology. General activities of acid and alkaline proteases were evaluated, as well as the effect of pH, temperature and inhibitors. The optimal pH of stomach proteases was 2, while the optimal pH of intestinal proteases was 10. Optimal temperature for the acidic proteases was 35 ºC, while for alkaline proteases it was 45 ºC. Thermal stability showed high resistance at 35 ºC for both acid and alkaline proteases (above 100% residual activity). Acid proteases are resistant at pH 2 (50% of residual activity), meanwhile alkaline proteases were highly resistant at pH 10 (90% of residual activity). Acid proteases were inhibited by 80% with pepstatin A and alkaline proteases were inhibited with TLCK and TPCK for trypsin (75%) and chymotrypsin (80%), respectively. Finally, metallo-proteases were 75% partially inhibited some serine proteases by 75% with EDTA. In conclusion, M. brachyurus has a good digestive capacity, since they can degrade a wide variety of proteins due to their greater proteolytic activity.

First molecular detection of Haemoproteus spp. and Plasmodium spp. in eared doves (Zenaida auriculata) in Brazil / Primeira detecção molecular de Haemoproteus spp. e Plasmodium spp. em pombos (Zenaida auriculata) no Brasil

The aim of this study was to verify the presence and identify the species of haemosporidian parasites in eared doves (Zenaida auriculata) in Brazil. Two hundred and eleven male and female eared doves were trap-captured in four different regions of Londrina city, in southern Brazil. Whole blood was collected in EDTA tubes through heart puncture after euthanasia in a CO2 chamber. A nested PCR targeting the mitochondrial cytochrome b gene (cyt b) of Haemoproteus spp./Plasmodium spp. was performed, followed by an enzymatic digestion to identify the genus. Phylogenetic trees were constructed to determine the closely related species. Out of 211 eared doves, 209 (99.05%) were positive for Haemoproteus spp. and/or Plasmodium spp. RFLP analysis showed that 72.72% (152/209) of eared doves were positive only for Haemoproteus spp., 6.22% (13/209) were positive only for Plasmodium spp., and 21.05% (44/209) of eared doves had mixed infections. Genetic analysis found four samples that were homologous with Haemoproteus multipigmentatus and one that was homologous with Plasmodium sp. This is the first molecular study of hemoparasites from eared doves in Brazil, and it is also the first description of H. multipigmentatus and Plasmodium spp. infection in eared doves in Brazil.(AU)

O objetivo deste estudo foi verificar a presença e a identificação espécies de parasitas hemosporídeos em pombos (Zenaida auriculata) no Brasil. Duzentos e onze pombos machos e fêmeas foram capturados em quatro regiões diferentes de Londrina, sul do Brasil. Amostra de sangue foi coletada em tubos contendo EDTA por meio de punção cardíaca, após eutanásia em câmara de CO2. Uma nested PCR com alvo no gene mitocondrial citocromo b (cyt b) de Haemoproteus spp./Plasmodium spp. foi realizada, seguida de digestão enzimática para identificar o gênero. A árvore filogenética foi construída para determinar a relação com outras espécies. Das 211 pombas, 209 (99,05%) foram positivas para Haemoproteus spp./Plasmodium spp. A análise RFLP demonstrou que 72,72% (152/209) das pombas foram positivas somente para Haemoproteus spp.; 6,22% (13/209) foram positivas somente para Plasmodium e 21,05% (44/209) das pombas tiveram infecções mistas. A análise genética mostrou quatro amostras homólogas com H. multipigmentatus e uma com Plasmodium spp. Este é o primeiro estudo molecular de hemoparasitas em pombos no Brasil. E é também a primeira descrição da infecção por H. multipigmentatus e Plasmodium spp. em pombos Z. auriculata no Brasil.(AU)

Partial characterization of digestive proteases in juveniles of Microphis brachyurus (short-tailed pipefish) (Syngnathiformes: Syngnathidae)

Short-tailed pipe fish (Microphis brachyurus) is a freshwater organism with high economic potential for the aquarium hobby, so it is necessary to implement methods to promote its culture through studies of digestive physiology. General activities of acid and alkaline proteases were evaluated, as well as the effect of pH, temperature and inhibitors. The optimal pH of stomach proteases was 2, while the optimal pH of intestinal proteases was 10. Optimal temperature for the acidic proteases was 35 ºC, while for alkaline proteases it was 45 ºC. Thermal stability showed high resistance at 35 ºC for both acid and alkaline proteases (above 100% residual activity). Acid proteases are resistant at pH 2 (50% of residual activity), meanwhile alkaline proteases were highly resistant at pH 10 (90% of residual activity). Acid proteases were inhibited by 80% with pepstatin A and alkaline proteases were inhibited with TLCK and TPCK for trypsin (75%) and chymotrypsin (80%), respectively. Finally, metallo-proteases were 75% partially inhibited some serine proteases by 75% with EDTA. In conclusion, M. brachyurus has a good digestive capacity, since they can degrade a wide variety of proteins due to their greater proteolytic activity.(AU)

El pez pipa (Microphis brachyurus) es un organismo dulceacuícola con alto potencial económico para la acuarofilia; sin embargo, es necesario implementar su cultivo a través de estudios de fisiología digestiva. Se evaluó el efecto del pH, temperatura e inhibidores sobre las actividades enzimáticas de proteasas ácidas y alcalinas. El pH óptimo de proteasas estomacales es de 2, mientras que el de proteases intestinales es de 10. La temperatura óptima de proteasas ácidas es de 35 ºC y las alcalinas de 45 ºC. La estabilidad térmica para proteasas ácidas y alcalinas es a los 35 ºC (más de 100% de actividad residual). La estabilidad a los diferentes pH de las proteasas ácidas es en 2 (50 % de la actividad residual), mientras que para las proteasas alcalinas es en 10 (90 % de la actividad residual). Las proteasas ácidas fueron inhibidas en 80% con pepstatina A y las proteasas alcalinas fueron altamente inhibidas con TLCK para tripsina (75%) y TPCK quimitripsina (80%). Finalmente, las metaloproteasas fueron inactivadas con EDTA en 70%. En conclusión, M. brachyurus tiene una buena capacidad digestiva al degradar una amplia variedad de proteinas debido a su alta actividad proteolítica.(AU)

The epiphyseal plate closure phenomena for male and female Japanese quail ( Coturnix coturnix japonica ): histological and biochemical alterations

This study was conducted to determine the effect of sex on ossification processes in quail by determining the exact closure timing of the epiphyseal plate in the proximal region of the femur. This was done by investigating the histological and biochemical parameters affecting the osteogenesis process that takes place following quail hatching to observe if any variation existed between males and females in this regard. For this purpose, blood samples were collected from six male and female specimens via IV catheters every week for the first 42 days that followed hatching. The samples were transferred into serum tubes, and PTH (PTH), triiodothyronine (T3), thyroxine (T4), inorganic phosphorus, calcium, and vitamin D values in the samples, which are known to have an impact on ossification, were analyzed. The specimens from which the blood samples were collected were then euthanized, and histological cut-sections that covered the epiphyseal growth plate were collected, along with the bone sections of the proximal regions of the right femur. Considering decalcification, these histological sections were kept in an ethylene diamine tetra acetic acid (EDTA) solution. Routine histological examinations were then conducted on these sections, after which they were embedded in paraffin. Crossman's modified triple staining method was used to prepare them, and Wilcoxon Signed Ranks Test was used to statistically evaluate whether the inspected biochemical parameters played a role in the ossification process of quail and whether a statistical difference existed between sexes in this regard. The findings of our study revealed that poultry animals also have five zones in the epiphyseal plate as do mammals, and they have calcified cartilage areas. The findings also indicate that, while ossification starts to occur both in male and female quail specimens, calcification occurs more frequently in females. It was determined that the proximal epiphyseal plate regions of the femur close at the end of the sixth week in both sexes of quail, although the calcification and ossification are more advanced in some females compared with males.(AU)